Palmitoylation of the cysteine-rich endodomain of the SARS-coronavirus spike glycoprotein is important for spike-mediated cell fusion.
Identifieur interne : 003643 ( Main/Exploration ); précédent : 003642; suivant : 003644Palmitoylation of the cysteine-rich endodomain of the SARS-coronavirus spike glycoprotein is important for spike-mediated cell fusion.
Auteurs : Chad M. Petit [États-Unis] ; Vladimir N. Chouljenko ; Arun Iyer ; Robin Colgrove ; Michael Farzan ; David M. Knipe ; K G KousoulasSource :
- Virology [ 0042-6822 ] ; 2007.
Descripteurs français
- KwdFr :
- Acide palmitique (analyse), Acide palmitique (métabolisme), Animaux, Cellules Vero, Cystéine (génétique), Cystéine (physiologie), Données de séquences moléculaires, Fusion cellulaire, Fusion membranaire, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (), Glycoprotéines membranaires (génétique), Glycoprotéines membranaires (métabolisme), Immunohistochimie, Marquage isotopique, Maturation post-traductionnelle des protéines, Membrane cellulaire (), Mutagenèse dirigée, Protéines de l'enveloppe virale (), Protéines de l'enveloppe virale (génétique), Protéines de l'enveloppe virale (métabolisme), Séquence d'acides aminés, Tritium (métabolisme), Virus du SRAS (), Virus du SRAS (physiologie).
- MESH :
- analyse : Acide palmitique.
- génétique : Cystéine, Glycoprotéines membranaires, Protéines de l'enveloppe virale.
- métabolisme : Acide palmitique, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Tritium.
- physiologie : Cystéine, Virus du SRAS.
- Animaux, Cellules Vero, Données de séquences moléculaires, Fusion cellulaire, Fusion membranaire, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires, Immunohistochimie, Marquage isotopique, Maturation post-traductionnelle des protéines, Membrane cellulaire, Mutagenèse dirigée, Protéines de l'enveloppe virale, Séquence d'acides aminés, Virus du SRAS.
English descriptors
- KwdEn :
- Amino Acid Sequence, Animals, Cell Fusion, Cell Membrane (chemistry), Chlorocebus aethiops, Cysteine (genetics), Cysteine (physiology), Immunohistochemistry, Isotope Labeling, Membrane Fusion, Membrane Glycoproteins (chemistry), Membrane Glycoproteins (genetics), Membrane Glycoproteins (metabolism), Molecular Sequence Data, Mutagenesis, Site-Directed, Palmitic Acid (analysis), Palmitic Acid (metabolism), Protein Processing, Post-Translational, SARS Virus (chemistry), SARS Virus (physiology), Spike Glycoprotein, Coronavirus, Tritium (metabolism), Vero Cells, Viral Envelope Proteins (chemistry), Viral Envelope Proteins (genetics), Viral Envelope Proteins (metabolism).
- MESH :
- chemical , analysis : Palmitic Acid.
- chemical , chemistry : Membrane Glycoproteins, Viral Envelope Proteins.
- chemical , genetics : Cysteine, Membrane Glycoproteins, Viral Envelope Proteins.
- chemistry : Cell Membrane, SARS Virus.
- chemical , metabolism : Membrane Glycoproteins, Palmitic Acid, Tritium, Viral Envelope Proteins.
- chemical , physiology : Cysteine, SARS Virus.
- Amino Acid Sequence, Animals, Cell Fusion, Chlorocebus aethiops, Immunohistochemistry, Isotope Labeling, Membrane Fusion, Molecular Sequence Data, Mutagenesis, Site-Directed, Protein Processing, Post-Translational, Spike Glycoprotein, Coronavirus, Vero Cells.
Abstract
The SARS-coronavirus (SARS-CoV) is the etiological agent of the severe acute respiratory syndrome (SARS). The SARS-CoV spike (S) glycoprotein mediates membrane fusion events during virus entry and virus-induced cell-to-cell fusion. The cytoplasmic portion of the S glycoprotein contains four cysteine-rich amino acid clusters. Individual cysteine clusters were altered via cysteine-to-alanine amino acid replacement and the modified S glycoproteins were tested for their transport to cell-surfaces and ability to cause cell fusion in transient transfection assays. Mutagenesis of the cysteine cluster I, located immediately proximal to the predicted transmembrane, domain did not appreciably reduce cell-surface expression, although S-mediated cell fusion was reduced by more than 50% in comparison to the wild-type S. Similarly, mutagenesis of the cysteine cluster II located adjacent to cluster I reduced S-mediated cell fusion by more than 60% compared to the wild-type S, while cell-surface expression was reduced by less than 20%. Mutagenesis of cysteine clusters III and IV did not appreciably affect S cell-surface expression or S-mediated cell fusion. The wild-type S was palmitoylated as evidenced by the efficient incorporation of (3)H-palmitic acid in wild-type S molecules. S glycoprotein palmitoylation was significantly reduced for mutant glycoproteins having cluster I and II cysteine changes, but was largely unaffected for cysteine cluster III and IV mutants. These results show that the S cytoplasmic domain is palmitoylated and that palmitoylation of the membrane proximal cysteine clusters I and II may be important for S-mediated cell fusion.
DOI: 10.1016/j.virol.2006.10.034
PubMed: 17134730
Affiliations:
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Le document en format XML
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence</term>
<term>Animals</term>
<term>Cell Fusion</term>
<term>Cell Membrane (chemistry)</term>
<term>Chlorocebus aethiops</term>
<term>Cysteine (genetics)</term>
<term>Cysteine (physiology)</term>
<term>Immunohistochemistry</term>
<term>Isotope Labeling</term>
<term>Membrane Fusion</term>
<term>Membrane Glycoproteins (chemistry)</term>
<term>Membrane Glycoproteins (genetics)</term>
<term>Membrane Glycoproteins (metabolism)</term>
<term>Molecular Sequence Data</term>
<term>Mutagenesis, Site-Directed</term>
<term>Palmitic Acid (analysis)</term>
<term>Palmitic Acid (metabolism)</term>
<term>Protein Processing, Post-Translational</term>
<term>SARS Virus (chemistry)</term>
<term>SARS Virus (physiology)</term>
<term>Spike Glycoprotein, Coronavirus</term>
<term>Tritium (metabolism)</term>
<term>Vero Cells</term>
<term>Viral Envelope Proteins (chemistry)</term>
<term>Viral Envelope Proteins (genetics)</term>
<term>Viral Envelope Proteins (metabolism)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Acide palmitique (analyse)</term>
<term>Acide palmitique (métabolisme)</term>
<term>Animaux</term>
<term>Cellules Vero</term>
<term>Cystéine (génétique)</term>
<term>Cystéine (physiologie)</term>
<term>Données de séquences moléculaires</term>
<term>Fusion cellulaire</term>
<term>Fusion membranaire</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Glycoprotéines membranaires ()</term>
<term>Glycoprotéines membranaires (génétique)</term>
<term>Glycoprotéines membranaires (métabolisme)</term>
<term>Immunohistochimie</term>
<term>Marquage isotopique</term>
<term>Maturation post-traductionnelle des protéines</term>
<term>Membrane cellulaire ()</term>
<term>Mutagenèse dirigée</term>
<term>Protéines de l'enveloppe virale ()</term>
<term>Protéines de l'enveloppe virale (génétique)</term>
<term>Protéines de l'enveloppe virale (métabolisme)</term>
<term>Séquence d'acides aminés</term>
<term>Tritium (métabolisme)</term>
<term>Virus du SRAS ()</term>
<term>Virus du SRAS (physiologie)</term>
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<keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Palmitic Acid</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Membrane Glycoproteins</term>
<term>Viral Envelope Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Cysteine</term>
<term>Membrane Glycoproteins</term>
<term>Viral Envelope Proteins</term>
</keywords>
<keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Acide palmitique</term>
</keywords>
<keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Cell Membrane</term>
<term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Cystéine</term>
<term>Glycoprotéines membranaires</term>
<term>Protéines de l'enveloppe virale</term>
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<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Membrane Glycoproteins</term>
<term>Palmitic Acid</term>
<term>Tritium</term>
<term>Viral Envelope Proteins</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Acide palmitique</term>
<term>Glycoprotéines membranaires</term>
<term>Protéines de l'enveloppe virale</term>
<term>Tritium</term>
</keywords>
<keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Cystéine</term>
<term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="physiology" xml:lang="en"><term>Cysteine</term>
<term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term>
<term>Animals</term>
<term>Cell Fusion</term>
<term>Chlorocebus aethiops</term>
<term>Immunohistochemistry</term>
<term>Isotope Labeling</term>
<term>Membrane Fusion</term>
<term>Molecular Sequence Data</term>
<term>Mutagenesis, Site-Directed</term>
<term>Protein Processing, Post-Translational</term>
<term>Spike Glycoprotein, Coronavirus</term>
<term>Vero Cells</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Animaux</term>
<term>Cellules Vero</term>
<term>Données de séquences moléculaires</term>
<term>Fusion cellulaire</term>
<term>Fusion membranaire</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Glycoprotéines membranaires</term>
<term>Immunohistochimie</term>
<term>Marquage isotopique</term>
<term>Maturation post-traductionnelle des protéines</term>
<term>Membrane cellulaire</term>
<term>Mutagenèse dirigée</term>
<term>Protéines de l'enveloppe virale</term>
<term>Séquence d'acides aminés</term>
<term>Virus du SRAS</term>
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<front><div type="abstract" xml:lang="en">The SARS-coronavirus (SARS-CoV) is the etiological agent of the severe acute respiratory syndrome (SARS). The SARS-CoV spike (S) glycoprotein mediates membrane fusion events during virus entry and virus-induced cell-to-cell fusion. The cytoplasmic portion of the S glycoprotein contains four cysteine-rich amino acid clusters. Individual cysteine clusters were altered via cysteine-to-alanine amino acid replacement and the modified S glycoproteins were tested for their transport to cell-surfaces and ability to cause cell fusion in transient transfection assays. Mutagenesis of the cysteine cluster I, located immediately proximal to the predicted transmembrane, domain did not appreciably reduce cell-surface expression, although S-mediated cell fusion was reduced by more than 50% in comparison to the wild-type S. Similarly, mutagenesis of the cysteine cluster II located adjacent to cluster I reduced S-mediated cell fusion by more than 60% compared to the wild-type S, while cell-surface expression was reduced by less than 20%. Mutagenesis of cysteine clusters III and IV did not appreciably affect S cell-surface expression or S-mediated cell fusion. The wild-type S was palmitoylated as evidenced by the efficient incorporation of (3)H-palmitic acid in wild-type S molecules. S glycoprotein palmitoylation was significantly reduced for mutant glycoproteins having cluster I and II cysteine changes, but was largely unaffected for cysteine cluster III and IV mutants. These results show that the S cytoplasmic domain is palmitoylated and that palmitoylation of the membrane proximal cysteine clusters I and II may be important for S-mediated cell fusion.</div>
</front>
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<tree><noCountry><name sortKey="Chouljenko, Vladimir N" sort="Chouljenko, Vladimir N" uniqKey="Chouljenko V" first="Vladimir N" last="Chouljenko">Vladimir N. Chouljenko</name>
<name sortKey="Colgrove, Robin" sort="Colgrove, Robin" uniqKey="Colgrove R" first="Robin" last="Colgrove">Robin Colgrove</name>
<name sortKey="Farzan, Michael" sort="Farzan, Michael" uniqKey="Farzan M" first="Michael" last="Farzan">Michael Farzan</name>
<name sortKey="Iyer, Arun" sort="Iyer, Arun" uniqKey="Iyer A" first="Arun" last="Iyer">Arun Iyer</name>
<name sortKey="Knipe, David M" sort="Knipe, David M" uniqKey="Knipe D" first="David M" last="Knipe">David M. Knipe</name>
<name sortKey="Kousoulas, K G" sort="Kousoulas, K G" uniqKey="Kousoulas K" first="K G" last="Kousoulas">K G Kousoulas</name>
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<country name="États-Unis"><noRegion><name sortKey="Petit, Chad M" sort="Petit, Chad M" uniqKey="Petit C" first="Chad M" last="Petit">Chad M. Petit</name>
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